Genome Editing in Citrus Tree with CRISPR/Cas9

  • Hongge Jia
  • Xiuping Zou
  • Vladimir Orbovic
  • Nian WangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)


CRISPR/Cas9 has been widely employed to edit genome in most of the organisms, including animal, plant, fungus, and microbe. Here we describe the modification of citrus gene CsLOB1 in transgenic citrus by Cas9/sgRNA, a two-component system derived from CRISPR-Cas9. Transgenic citrus plants can be created by Agrobacterium-mediated epicotyl transformation.

Key words

Cas9/sgRNA Genome editing Citrus Agrobacterium-mediated epicotyl transformation CsLOB1 



This work has been supported by Florida Citrus Initiative.


  1. 1.
    Hsu PD, Lander ES, Zhang F (2014) Development and applications of CRISPR-Cas9 for genome engineering. Cell 157:1262–1278CrossRefGoogle Scholar
  2. 2.
    Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821CrossRefGoogle Scholar
  3. 3.
    Jia H, Wang N (2014a) Targeted genome editing of sweet orange using Cas9/sgRNA. PLoS One 9:e93806CrossRefGoogle Scholar
  4. 4.
    Jia H, Wang N (2014b) Xcc-facilitated agroinfiltration of citrus leaves: a tool for rapid functional analysis of transgenes in citrus leaves. Plant Cell Rep 33:1993–2001CrossRefGoogle Scholar
  5. 5.
    Jia H, Orbovic V, Jones JB, Wang N (2016) Modification of the PthA4 effector binding elements in Type I CsLOB1 promoter using Cas9/sgRNA to produce transgenic Duncan grapefruit alleviating XccΔpthA4:dCsLOB1.3 infection. Plant Biotechnol J 14:1291–1301CrossRefGoogle Scholar
  6. 6.
    Jia H, Zhang Y, Orbovic V, Xu J, White FF, Jones JB, Wang N (2017) Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker. Plant Biotechnol J 15(7):817–823CrossRefGoogle Scholar
  7. 7.
    Peng A, Chen S, Lei T, Xu L, He Y, Wu L, Yao L, Zou X (2017) Engineering canker-resistant plants through CRISPR/Cas9-targeted editing of the susceptibility gene CsLOB1 promoter in citrus. Plant Biotechnol J 15:1509. Scholar
  8. 8.
    Mao Y, Botella JR Zhu JK (2017) Heritability of targeted gene modifications induced by plant-optimized CRISPR systems. Cell Mol Life Sci 74(6):1075–1093CrossRefGoogle Scholar
  9. 9.
    Liu X, Xie C, Si H, Yang J (2017) CRISPR/Cas9-mediated genome editing in plants. Methods 121-122:94–102CrossRefGoogle Scholar
  10. 10.
    Liu X, Wu S, X J, Sui C, Wei J (2017) Application of CRISPR/Cas9 in plant biology. Acta Pharm Sin B 7:292–302CrossRefGoogle Scholar
  11. 11.
    Malzahn A, Lowder L, Qi Y (2017) Plant genome editing with TALEN and CRISPR. Cell Biosci 7:21CrossRefGoogle Scholar
  12. 12.
    Demirci Y, Zhang B, Unver T (2017) CRISPR/Cas9: an RNA-guided highly precise synthetic tool for plant genome editing. J Cell Physiol 233:1844. Scholar
  13. 13.
    Donmez D, Simsek O, Izgu T, Kacar YA, Mendi YY (2013) Genetic transformation in citrus. Sci World J 2013:491207CrossRefGoogle Scholar
  14. 14.
    Jiang W, Yang B, Weeks DP (2014) Efficient CRISPR/Cas9-mediated gene editing in Arabidopsis thaliana and inheritance of modified genes in the T2 and T3 generations. PLoS One 9:e99225CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Hongge Jia
    • 1
  • Xiuping Zou
    • 1
  • Vladimir Orbovic
    • 2
  • Nian Wang
    • 1
    Email author
  1. 1.Citrus Research and Education Center, Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences (IFAS)University of FloridaLake AlfredUSA
  2. 2.Citrus Research and Education Center, Institute of Food and Agricultural Sciences (IFAS)University of FloridaLake AlfredUSA

Personalised recommendations